雌马酚对非酒精性脂肪性肝细胞模型的氧化应激作用研究  

Effect of equol on oxidative stress in a non-alcoholic fatty liver cell model

作　　者：崔涵强 倪向敏 张贵明 徐喆 李硕 王建 CUI Hanqiang;NI Xiangmin;ZHANG Guiming;XU Zhe;LI Shuo;WANG Jian(Department of Nutrition,the Second Affiliated Hospital of Army Medical University,Chongqing 400037,China)

机构地区：[1]陆军军医大学第二附属医院营养科,重庆400037

出　　处：《重庆医学》2023年第15期2241-2246,共6页Chongqing medicine

基　　金：国家自然科学基金面上项目(81973040);国家自然科学基金青年科学基金项目(81602855)。

摘　　要：目的观察雌马酚(Eq)对高脂诱导的非酒精性脂肪性肝病(NAFLD)细胞模型中氧化应激的保护效应,并探讨其作用机制。方法采用CCK-8法探索合适的油酸钠(NaOL)和Eq的干预浓度。通过NaOL作用于BRL3A细胞,建立NAFLD体外细胞模型,设立对照组(Con组)、模型组(Mod组)、低剂量组(Eq-L组)、中剂量组(Eq-M组)和高剂量组(Eq-H组)。干预24 h后,采用尼罗红荧光染色观察细胞内脂滴累积情况,检测细胞内甘油三脂(TG)、总胆固醇(TC)、活性氧(ROS)、超氧化物歧化酶(SOD)、丙二醛(MDA)和过氧化氢酶(CAT)的水平,采用蛋白免疫印迹和实时荧光定量聚合酶链式反应(qRT-PCR)检测细胞内Nrf2、HO-1、NQO1蛋白和mRNA的表达水平。结果与Con组比较,Mod组细胞内脂质累积增多,TG、TC、ROS、MDA水平升高,SOD、CAT水平降低,Nrf2、HO-1、NQO1蛋白和mRNA表达下调,差异均有统计学意义(P<0.05)。与Mod组比较,Eq-L组、Eq-M组和Eq-H组细胞内脂质累积逐渐减少,TG、TC、ROS、MDA水平降低,SOD、CAT水平增多,Nrf2、HO-1、NQO1蛋白和mRNA表达上调,差异均有统计学意义(P<0.05)。结论Eq通过抑制氧化应激反应改善NAFLD病情,其部分机制可能与Eq调控Nrf2信号通路相关。Objective To investigate the protective effect of Equol(Eq)on oxidative stress in a cell model of high-fat-induced non-alcoholic fatty liver disease(NAFLD),and to explore its mechanism.Methods The optimal concentration of sodium oleate(NaOL)and Eq for intervention was determined using the CCK-8 method.The in vitro cell model of NAFLD was established by treating BRL3A cells with NaOL.A control group(Con group),a model group(Mod group),a low-dose group(Eq-L group),a middle-dose group(Eq-M group),and a high-dose group(Eq-H group)were established.After 24 hours of intervention,Nile red fluorescent staining was used to observe the accumulation of intracellular lipid droplets and detect the levels of intracellular triglycerides(TG),total cholesterol(TC),reactive oxygen species(ROS),superoxide dismutase(SOD),malondialdehyde(MDA),and catalase(CAT).Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR)were used to detect the expression levels of nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),and NQO1 proteins and mRNAs in cells.Results Compared to the Con group,the Mod group showed an increase in intracellular lipid accumulation.Additionally,the levels of TG,TC,ROS,and MDA increased,while the levels of SOD and CAT decreased.Furthermore,the protein and mRNA expressions of Nrf2,HO-1,and NQO1 were down-regulated.These differences were statistically significant(P<0.05).Compared to the Mod group,the intracellular lipid accumulation in the Eq-L group,Eq-M group,and Eq-H group decreased gradually.The levels of TG,TC,ROS,and MDA also decreased,while the levels of SOD and CAT increased.Additionally,the expressions of Nrf2,HO-1,NQO1 protein,and mRNA were up-regulated,and these differences were statistically significant(P<0.05).Conclusion Eq improves the condition of NAFLD by inhibiting the oxidative stress response,and part of the mechanism may be related to the regulation of the Nrf2 signaling pathway by Eq.

关 键 词：雌马酚 氧化应激 非酒精性脂肪肝 核因子E2相关因子 蛋白质印迹法 

分 类 号：R575[医药卫生—消化系统]